Use of oligosaccharide for preventing blood clotting in extracorporeal blood circuits

ABSTRACT

A method for preventing clotting in an extracorporeal blood circuit by administering a synthetic oligosaccharide that is a selective inhibitor of factor Xa, acting via antithrobmin III.

The invention relates to the use of a certain oligosaccharide for themanufacture of a medicament for preventing blood clotting inextracorporeal blood circuits. Further the invention relates to apharmeuceutical composition for said use.

Blood clotting in extracorporeal blood circuits needs to be prevented.Otherwise, blood coagulation occurs as soon as blood contacts artificialsurfaces. As a remedy, usually unfractionated heparin (UFH) or lowmolecular weight heparins (LMWH) are used as anticoagulants.

Both UFH and LMWH have an effect on several stages of the bloodcoagulation cascade, both inhibiting factor Xa and thrombin (factorIIa). Factor Xa catalyzes the generation of thrombin and subsequentlythrombin regulates the last step in the coagulation cascade. The primefunction of thrombin is the cleavage of fibrinogen to generate fibrinmonomers, which form an insoluble gel by cross-linking, therebyinitiating thrombus formation. UFH and LMWH have thrombolyticproperties, i.e. they induce dissolution of the thrombus formed.

Contrary to UFH and LMWH, some synthetic oligosaccharides, especiallyoligosaccharides described in EP 84,899 and U.S. Pat. No. 5,378,829,highly selectively inhibit factor Xa via antithrombin III (ATIII) but donot have any activity on thrombin. However, notwithstanding the absenceof any capacity to inhibit thrombin or to promote thrombolysis, itappeared that those oligosaccharides inhibit thrombous formation, e.g.as occurring in extracorporeal blood circuits. Thus, surprisingly, ithas now been found that a synthetic oligosaccharide which is a selectiveinhibitor of factor Xa, acting via antithrobmin III, is useful forpreventing blood clotting in patients with an extracorporeal bloodcircuit.

The use of the oligosaccharide according to this invention results ineffective and safe inhibition of blood clotting, e.g. in patientsundergoing haemodialysis, without increased bleeding risks.

A preferred oligosaccharide for the use according to this invention isthe pentasaccharide with the formula methylO-(2-deoxy-2-sulphoamino-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(β-D-glucopyranosyl uronicacid)-(1→4)-O-(2-deoxy-2-sulphoamino-3,6-di-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2-O-sulpho-α-L-idopyranosyluronic acid)-(1→4)-2-deoxy-2-sulphoamino-6-O-sulpho-α-D-glucopyranosideor a pharmaceutically acceptable salt thereof (i.e. salts withcounter-ions like hydrogen or, more preferably, alkali or earth-alkalimetal ions, like sodium, calcium, or magnesium), having the structure:

Particularly preferred is its decasodium salt, known by its code nameOrg 31540 or SR 90107A (described in chemical Synthesis toGlycosaminoglycans, Supplement to Nature 1991, 350, 30-33).

Other advantageous pentasaccharides are: methylO-(3,4-di-O-methyl-2,6-di-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(3-O-methyl-2-O-sulpho-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(3-methyl-2-O-sulpho-α-L-idopyranosyluronic acid)-(1→4)-2,3,6-tri-O-sulpho-α-D-glucopyranoside or apharmaceutically acceptable salt thereof (especially its dodecasodiumsalt described in U.S. Pat. No. 5,378,829), having the structure

and methylO-(2,3,4-tri-O-methyl-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1→4)-2,3,6-tri-O-sulpho-α-D-glucopyranoside or apharmaceutically acceptable salt thereof (especially its nonasodium saltalso described in U.S. Pat. No. 5,378,829), having the structure

The use in patients with extracorporeal blood circuits according to theinvention includes circuits and intravenous infusion lines used forhaemodialysis, renal dialysis, haemofiltration, and the like. Preferredextracorporeal cicuits are those used in the treatment of haemodialysispatients.

The oligosaccharide can be administered at several stages of thetreatment. Preferably, but not limited to this route of administration,the oligosaccharide is administered as an intravenous injection to themammal undergoing treatment. Preferably, the mammal is a human.

Another route of administration of the oligosaccharide is theintroduction thereof into a (dialysis) circuit by other means, e.g. byinjecting it either gradually or at once into the system upstream of thedialysis membrane simultaneously with the introduction of the blood intothe circuit Moreover, the lines and/or further equipment of theextracorporeal circuit can be furnished with the oligosaccharide,preferably by way of a coating (but not limited to this). Alternatively,the oligosaccharide may be adsorbed in the materials of parts of theequipment, e.g. in the membranes used for dialysis.

For use according to the invention, the oligosaccharide may beadministered enterally or parenterally (especially via the subcutaneousor intravenous route) or may be administered via an external source(vide supra), and for humans preferably in a dosage of 0,001-10 mg perkg body weight per dialysis. More preferably, the pentasaccharide isadministered at doses of between 0.30 mg and 30 mg per patient perdialysis.

The oligosaccharide may be used alone or may be presented as apharmaceutical composition. Accordingly, the present invention furtherprovides a pharmaceutical composition for preventing blood clotting inan extracorporeal blood circuit comprising said oligosaccharide togetherwith pharmaceutically acceptable auxiliaries and optionally othertherapeutic agents. The term “acceptable” means being compatible withthe other ingredients of the composition and not deleterious to therecipients thereof.

Compositions include e.g. those suitable for oral, sublingual,subcutaneous, intravenous, intramuscular, transdermal, transmucosal,local, or rectal administration, and the like, all in unit dosage formsfor administration.

For oral administration, the active ingredient may be presented asdiscrete units, such as tablets, capsules, powders, granulates,solutions, suspensions, and the like. For parenteral administration, thepharmaceutical composition of the invention may be presented inunit-dose or multi-dose containers, e.g. injection liquids inpredetermined amounts, for example in sealed vials and ampoules, and mayalso be stored in a freeze dried (lyophilzed) condition requiring onlythe addition of sterile liquid carrier, e.g. water, prior to use.

Mixed with such pharmaceutically acceptable auxiliaries, e.g. asdescribed in the standard reference, Gennaro et al., Remington'sPharmaceutical Sciences, (18th ed., Mack Publishing Company, 1990, seeespecially Part 8: Pharmaceutical Preparations and Their Manufacture),the oligosaccharide may be compressed into solid dosage units, such aspills, tablets, or be processed into capsules or suppositories. By meansof pharmaceutically acceptable liquids the oligosaccharide can beapplied as a fluid composition, e.g. as an injection preparation, in theform of a solution, suspension, emulsion, or as a spray, e.g. a nasalspray.

For use as a coating according to the invention, for examplepharmaceutically acceptable polymers may be used as a matrix for theoligosaccharide. Also coatings are included, in which theoligosaccharide is chemically (e.g. covalently) linked to the surfacewithout loss of its activity. Any parmaceutically acceptable coating maybe suitable for this purpose, prepared according to methods conventionalin the art.

For making solid dosage units, the use of conventional additives such asfillers, colorants, polymeric binders and the like is contemplated. Ingeneral any pharmaceutically acceptable additive which does notinterfere with the function of the active compounds can be used.Suitable carriers with which the oligosaccharides of the invention canbe administered as solid compositions include lactose, starch, cellulosederivatives and the like, or mixtures thereof, used in suitable amounts.For parenteral administration, aqueous suspensions, isotone salinesolutions and sterile injectable solutions may be used, containingpharmaceutically acceptable dispersing agents and/or wetting agents,such as propylene glycol or butylene glycol.

The pharmaceutical composition according to the invention may also bepresented in the form of a veterinary composition, such compositions maybe prepared by methods conventional in the art.

The invention further includes a pharmaceutical composition, ashereinbefore described, in combination with packaging material suitablefor said composition, said packaging material including instructions forthe use of the composition for the use as hereinbefore described.

The invention is further illustrated by the following example. Thisshould not be considered to be limiting in any way.

EXAMPLE

The pentasaccharide Org 31540/SR 90107 A, as a representative compoundfor use according to the present invention, has been subject to a pilotclinical study in 12 patients undergoing chronic intermittenthaemodialysis.

The study consisted of 2 phases (block A and block B). In block A, 10 mgof Org 31540/SR 90107 A was administered. Thereafter, in block B, 8, 6,and 4 mg of Org 31540/SR 90107 A were used.

Medication was given as an intravenous bolus predialysis for 1 dialysiseach week. Efficacy was assessed by determining patency of the dialyzer,buffer and bubble chamber, every hour during dialysis by visualexamination and by blood sampling of specific coagualtion, heamatologicand biochemical parameters. Anti-Xa plasma samples, to determinepharmacokinetics, were taken every hour during dialysis and 1 hour afterdialysis and daily for 3 days post-dialysis. Safety was assessed byevaluating major and minor bleeding complications each dialysis.

Results: All patients have completed the study. Dialysis could beperformed without total clotting of the extracorporeal circuit in allpatients for all study dialyses. Only in one patient a clot in thebuffer chamber made dialysis impossible half an hour before the end ofthe last dialysis. No minor or major bleedings were recorded.

Conclusion: The pentasaccharide Org 31540/SR 90107 A is a safe (noincreased haemorrhagic risks) and effective (at several doses)anticoagulant to prevent clotting in the extracorporeal blood circuit inhaemodialysis patients.

1. A method for preventing clotting in an extracorporeal blood circuit,induced by contact with surfaces, for a patient undergoing chronic,intermittent, extracorporeal blood treatment, comprising: administeringto the patient for each treatment an amount of from 0.001 to 10 mg ofmethylO-(3,4-di-O-methyl-2,6-di-O-sulpho-α-D-glucopyranosyl-(1→4)-O-(3-O-methyl-2-O-sulpho-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,36-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(3-O-methyl-2-O-sulpho-α-L-idopyranosyluronic acid)-(1→4)-(2,3,6-tri-O-sulpho-α-D-glucopyranoside, or a saltthereof, per kg body weight of the patient.
 2. A method for preventingclotting in an extracorporeal blood circuit, induced by contact withsurfaces, for a patient undergoing chronic, intermittent, extracorporealblood treatment, comprising: administering to the patient for eachtreatment an amount of from 0.30 to 30 mg of methylO-(3,4-di-O-methyl-2,6-di-O-sulpho-α-D-glucopyranosyl-(1→4)-O-(3-O-methyl-2-O-sulpho-β-D-glucopyranosyluronicacid)-(1→4)-O--2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(3-O-methyl-2-O-sulpho-α-L-idopyranosyluronic acid)-(1→4)-2,3,6-tri-O-sulpho-α-D-glucopyranoside, or saltthereof.
 3. The method of claim 1, comprising administering adodecasodium salt thereof.
 4. The method of claim 2, comprisingadministering a dodecasodium salt thereof.
 5. A method for preventingclotting in an extracorporeal blood circuit, induced by contact withsurfaces, for a patient undergoing chronic, intermittent, extracorporealblood treatment, comprising: administering to the patient for eachtreatment an amount of from 0.001 to 10 mg of methylO-(2,3,4-tri-O-methyl-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1→4)-2,3,6,-tri-O-sulpho-α-D-glucopyranoside, or a saltthereof, per kg body weight of the patient.
 6. A method for preventingclotting in an extracorporeal blood circuit, induced by contact withsurfaces, for a patient undergoing chronic, intermittent, extracorporealblood treatment, comprising: administering to the patient for eachtreatment an amount of from 0.30 to 30 mg of methylO-(2,3,4-tri-O-methyl-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1→4)-2,3,6,-tri-O-sulpho-α-D-glucopyranoside, or a saltthereof.
 7. The method of claim 5, comprising administering a nonasodiumsalt thereof.
 8. The method of claim 6, comprising administering anonasodium salt thereof.
 9. A method for preventing clotting in anextracorporeal blood circuit, induced by contact with surfaces, for apatient undergoing chronic, intermittent, extracorporeal bloodtreatment, comprising: administering to the circuit for each treatmentan amount form 0.001 to 10 mg of methylO-(3,4-di-O-methyl-2,6-di-O-sulpho-α-D-glucopyranosyl-(1→4)-O-(3-O-methyl-2-O-sulpho-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(3-O-methyl-2-O-sulpho-α-L-idipyranosyluronic acid)-(1→4)-2,3,6-tri-O-sulpho-α-D-glucopyranoside, or saltthereof, per kg body weight of the patient.
 10. A method for preventingclotting in an extracorporeal blood circuit, induced by contact withsurfaces, for a patient undergoing chronic, intermittent, extracorporealblood treatment, comprising: administering to the circuit for eachtreatment an amount of from 0.30 to 30 mg of methylO-(3,4-di-O-methyl-2,6-di-O-sulpho-α-D-glucopyranosyl-(1→4)-O-(3-O-methyl-2-O-sulpho-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(→4)-O-(3-O-methyl-2-O-sulpho-α-L-idopyranosyluronic acid)-(1→4)-(2,3,6-tri-O-sulpho-α-D-glucopyranoside or a saltthereof.
 11. The method of claim 9, comprising administering adodecasodium salt thereof.
 12. The method of claim 10, comprisingadministering a dodecasodium salt thereof.
 13. A method for preventingclotting in an extracorporeal blood circuit, induced by contact withsurfaces, for a patient undergoing chronic, intermittent, extracorporealblood treatment, comprising: administering to the circuit for eachtreatment an amount of from 0.001 to 10 mg of methylO-(2,3,4-tri-O-methyl-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1→4)-2,3,6-tri-O-sulpho-α-D-glucopyranoside or a saltthereof per kg body weight of the patient.
 14. A method for preventingclotting induced by contact with surfaces in an extracorporeal bloodcircuit for a patient undergoing chronic, intermittent, extracorporealblood treatment comprising: administering to the circuit for eachtreatment an amount of from 0.30 to 30 mg of methylO-(2,3,4-tri-O-methyl-6-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-β-D-glucopyranosyluronicacid)-(1→4)-O-(2,3,6-tri-O-sulpho-α-D-glucopyranosyl)-(1→4)-O-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1→4)-2,3,6,-tri-O-sulpho-α-D-glucopyranoside or a saltthereof.
 15. The method of claim 13, comprising administering anonasodium salt thereof.
 16. The method of claim 14, comprisingadministering a nonasodium salt thereof.